黄连解毒汤对急性鼻窦炎模型大鼠鼻窦黏膜MAPK/NF-κB信号通路的影响

Effect of Huanglian Jiedu Decoction(黄连解毒汤)on MAPK/NF-κB Signaling Pathway in Sinus Mucosa of Rats with Acute Sinusitis

目的:探讨黄连解毒汤对急性鼻窦炎模型大鼠鼻窦黏膜丝裂原活化蛋白激酶(MAPK)/核因子-κB(NF-κB)信号通路的影响。方法:69只SD大鼠随机取10只作为空白对照组,其余59只SD大鼠采用肺炎链球菌混合膨胀海绵鼻腔填塞方法建立急性鼻窦炎模型,造模成功后,将50只SD大鼠随机分为模型组、黄连解毒汤低剂量组、黄连解毒汤中剂量组、黄连解毒汤高剂量组和阳性对照组,每组10只。各给药组大鼠采用鼻腔灌洗的方式给予相应药物,模型组和空白对照组大鼠给予等体积生理盐水鼻腔灌洗,连续给药7 d。给药结束后,观察大鼠体征并进行鼻窦炎症状评分;鼻腔灌洗液培养观察肺炎链球菌生长情况;苏木精-伊红(HE)染色观察大鼠鼻窦黏膜病理损伤;酶联免疫吸附法(ELISA)检测血清中炎症因子(TNF-α、IL-6)含量;qRT-PCR检测鼻窦黏膜TNF-αmRNA、IL-6 mRNA水平;Western bloting检测鼻窦黏膜TNF-α、IL-6及C-Jun N末端激酶(JNK)、细胞外信号调节激酶1/2(ERK1/2)、p38、NF-κB p65水平。结果:与空白对照组比较,模型组大鼠鼻分泌物的细菌CFU值明显升高,鼻窦黏膜有明显的炎症反应,血清IL-6和TNF-α水平明显升高,鼻窦黏膜中JNK、ERK1/2、p38、NF-κB p65磷酸化水平明显升高,差异均有统计学意义(P<0.05)。与模型组比较,黄连解毒汤高、中、低剂量组和阳性对照组大鼠鼻分泌物的细菌CFU值明显降低,鼻窦黏膜的炎症反应明显改善,血清IL-6和TNF-α水平明显降低,鼻窦黏膜中JNK、ERK1/2、p38、NF-κB p65磷酸化水平明显降低,差异均有统计学意义(P<0.05)。结论:黄连解毒汤对大鼠鼻窦黏膜具有剂量依赖的保护作用,其可能与抑制MAPK/NF-κB信号通路的激活,以剂量依赖的方式下调炎症因子的基因转录,从而减少炎症因子的产生有关。

Objective:To investigate the effect of Huanglian Jiedu Decoction on mitogen-activated protein kinase(MAPK)/nuclear factor-κB(NF-κB)signaling pathway in sinus mucosa of rats with acute sinusitis.Methods:10 of 69 SD rats were randomly selected as the blank control group,and the remaining 59 SD rats were used to establish an acute sinusitis model by nasal packing with Streptococcus pneumoniae mixed with expanded sponge.After successful modeling,50 SD rats were randomly divided into model group,Huanglian Jiedu Decoction low-dose group,Huanglian Jiedu Decoction medium-dose group,Huanglian Jiedu Decoction high-dose group and positive control group,with 10 rats in each group.Rats in each administration group were given corresponding drugs by nasal lavage,while rats in model group and blank control group were given equal volume of normal saline by nasal lavage for 7 d.After the administration,the physical signs of the rats were observed and the symptoms of sinusitis were scored.The growth of Streptococcus pneumoniae was observed by nasal lavage fluid culture.Hematoxylin eosin(HE)staining was used to observe the pathological damage of sinus mucosa in rats.The contents of inflammatory factor(TNF-α,IL-6)in serum by enzyme-linked immunosorbent assay(ELISA).The levels of TNF-αmRNA and IL-6 mRNA in sinus mucosa were detected by qRT-PCR.The levels of TNF-α,IL-6,c-Jun N-terminal kinase(JNK),extracellular signal-regulated kinase 1/2(ERK1/2),p38 and NF-κB p65 were detected by Western blotting.Results:Compared with the blank control group,the bacterial CFU value of nasal secretions in the model group was significantly increased,the sinus mucosa had obvious inflammatory response,the levels of serum IL-6 and TNF-αwere significantly increased,and the phosphorylation levels of JNK,ERK1/2,p38 and NF-κB p65 in the sinus mucosa were significantly increased(P<0.05).Compared with the model group,the bacterial CFU value of nasal secretions of the rats in the Huanglian Jiedu Decoction low,medium and high-dose groups and the positive control group were obviously decreased,the inflammatory response of the sinus mucosa was obviously improved,the levels of serum IL-6 and TNF-αwere significantly decreased,and the phosphorylation levels of JNK,ERK1/2,p38 and NF-κB p65 in sinus mucosa were significantly decreased(P<0.05).Conclusion:Huanglian Jiedu Decoction has a dose-dependent protective effect on rat sinus mucosa,which may be related to inhibiting the activation of MAPK/NF-κB signaling pathway and down-regulating the gene transcription of inflammatory factors in a dose-dependent manner,thereby reducing the production of inflammatory factors.